stable cell lines lentiviral particles  (Cell Biolabs Inc)

Journal: Cell Death & Disease

Article Title: MCL1 inhibition targets Myeloid Derived Suppressors Cells, promotes antitumor immunity and enhances the efficacy of immune checkpoint blockade

doi: 10.1038/s41419-024-06524-w

Figure Lengend Snippet: A KM plot (kmplot.com) created from transcriptomic dataset of cancer patients treated with Anti-PD-1 based on the expression of MCL1. The p -value and FDR value are listed in the graph. B Combination treatment schema. C Tumor volumes in the C57BL/6J syngeneic mouse model with melanoma line B16.F10. The combination treatment of S64315 with anti-PD-1 significantly inhibited the tumor growth, compared to vehicle or the single drugs for multiple days. For visual clarification, we marked only the last day. ** p < 0.01. Within each significant treatment, the least significant p -value of the comparisons is displayed. Error bars represent ±SEM for all figures.

Article Snippet: For mouse studies with MCL1 knock down lines, B16.F10 and YUMM1.7 sh Control or sh MCL1 stable cell lines were created using shRNA lentiviral particles (Santa Cruz Biotechnology, Dallas, TX, USA).

Techniques: Expressing, Clarification Assay

Journal: Cell Death & Disease

Article Title: MCL1 inhibition targets Myeloid Derived Suppressors Cells, promotes antitumor immunity and enhances the efficacy of immune checkpoint blockade

doi: 10.1038/s41419-024-06524-w

Figure Lengend Snippet: Effects of the MCL1 inhibitor S64315 on growth of B16.F10 tumors in immunocompromised nude mice ( A ) or in immunocompetent C57BL/6 J mice ( B ). Day 0 was the day of cell implantation. Treatments were 25 mg/kg, at the Regimen 1 (Days 4, 6, 11, and 13) or Regimen 2 (Days 4, 6, 9, 11, and 13). C Effects of S64315 on B16.F10 and YUMM1.7 cells in vitro. Y-axis shows the relative viability compared to the cells treated with the vehicle control, and dashed line indicates 50% viability. D Effects of MCL1 knockdown of B16.F10 and YUMM1.7 on their tumor growth, with cells of the control (sh Control) and knockdown (sh MCL1). *** p < 0.001. Error bars represent ±SEM for all figures. Immunoblot of the knockdown are in Supplementary Fig. .

Article Snippet: For mouse studies with MCL1 knock down lines, B16.F10 and YUMM1.7 sh Control or sh MCL1 stable cell lines were created using shRNA lentiviral particles (Santa Cruz Biotechnology, Dallas, TX, USA).

Techniques: In Vitro, Control, Knockdown, Western Blot

Journal: Cell Death & Disease

Article Title: MCL1 inhibition targets Myeloid Derived Suppressors Cells, promotes antitumor immunity and enhances the efficacy of immune checkpoint blockade

doi: 10.1038/s41419-024-06524-w

Figure Lengend Snippet: Human MDSCs and T-cells were generated from five separate healthy donors. A Viability assays with human MDSCs, human melanoma cell line A375, mouse melanoma line B16.F10, and human MDSC-like cell line HL-60 after 24 h treatments with various doses of the MCL1 inhibitor S64315 in vitro. The bottom panel shows the IC50 values. B Quantification of the CD8+T cell proliferation in PBMCs activation with anti-CD3/CD28 beads analyzed at 7 days post activation. Example histogram of flow cytometric analysis of CD8+T cell proliferation. C Quantification of the CD4+T cell proliferation in PBMC activation with anti-CD3/CD28 beads, analyzed at 7 days post activation. Example histogram of flow cytometric analysis of CD4+T cell proliferation. D Quantification of intracellular cytokine staining for CD8+T cell expression of IFNγ and GZMB at 7 days post activation. E Representative contour plots showing the expression of IFNγ and GZMB in CD8+T cells. *** p < 0.001. Y -axis shows percentage of relative viability and X -axis indicates the dosages of the MCL1 inhibitor S64315 in µM. The dashed line indicates 50% viability. Error bars represent ±SEM for all figures.

Article Snippet: For mouse studies with MCL1 knock down lines, B16.F10 and YUMM1.7 sh Control or sh MCL1 stable cell lines were created using shRNA lentiviral particles (Santa Cruz Biotechnology, Dallas, TX, USA).

Techniques: Generated, In Vitro, Activation Assay, Staining, Expressing

Journal: Cell Death & Disease

Article Title: MCL1 inhibition targets Myeloid Derived Suppressors Cells, promotes antitumor immunity and enhances the efficacy of immune checkpoint blockade

doi: 10.1038/s41419-024-06524-w

Figure Lengend Snippet: A UMAP projection of live/CD45+/CD3-/CD19-/CD56-/CD11b+ cells from one melanoma tumor (omental metastasis from 30-year-old patient that experienced progressive disease following 17 cycles of pembrolizumab). B Dot plot depicting the top 10 unique genes that define each of the clusters. C Dot plot depicting genes associated with the MDSC transcriptional profile. D Dot plot depicting the expression of genes in the BCL2 family. E Feature plots showing the distribution of the relative expression of MCL1, BCL2, and BCL2L1 respectively, as clustered in A . F Representative flow cytometric gating strategy for the analysis of MCL1, BCL2, and BCLXL expression in MDSCs from melanoma patients. G Representative histogram of flow cytometric analysis of MCL1, BCL2, and BCLXL expression induced by the labeled cytokines in MDSCs, gated as shown in F . Data in G are representative graphs from 10 separate melanoma patients (Supplementary Fig. ), comparing the labeled fluorochrome conjugated antibody to the corresponding fluorescence minus one (FMO) staining control. Y-axis values are normalized to the cell count in each sample. X-axis label includes antibody specificity and fluorochrome conjugate. # indicates cells expressing the MDSC transcriptional profile.

Article Snippet: For mouse studies with MCL1 knock down lines, B16.F10 and YUMM1.7 sh Control or sh MCL1 stable cell lines were created using shRNA lentiviral particles (Santa Cruz Biotechnology, Dallas, TX, USA).

Techniques: Expressing, Labeling, Fluorescence, Staining, Control, Cell Counting